Microperoxidases catalytically degrade reactive oxygen species and may be anti-cataract agents.
microPx-11, a ferriheme undecapeptide proteolytic degradation product of cytochrome C is shown to be a peroxidase with broad specificity degrading H2O2 and tertiary butyl hydroperoxide. It is also capable of effectively eliminating superoxide and hydroxyl radical. The peroxidase loses activity in the presence of peroxide unless it is stabilized by ascorbate (Asc) or solutions such as aqueous humor or medium 199. While thiol but not disulfides inactivates the microPx-11, it is not inhibited in the presence of the rat lens which has a high GSH content. microPx-11 at concentrations 10 to 50 fold greater than are required to achieve good protective activity exhibits no toxicity based on cell viability, ATP levels and lens transparency after long-term incubations of alpha TN4-1 cells or cultured rat lens. The peroxidase is capable of protecting cultured rat lenses from photochemical stress where H2O2, O2.- and OH. are generated based on transparency, choline transport, epithelial cell viability and protein integrity as indicated by SDS-PAGE of the rat lens protein. In the absence of the peroxidase, extensive epithelial cell death and other degradative changes are observed. The DNA of alpha TN4-1 cells can also be protected from H2O2 induced single strand breaks by the microPx-11. The overall results suggest that a number of cytochrome C proteolytic degradation products are peroxidases which may be effective anti-cataract agents protecting the lens from oxidative stress.
Spector
A
A
Department of Ophthalmology, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
Ma
W
W
Wang
R R
RR
Kleiman
N J
NJ
D000818:Animals; D002386:Cataract / Q000188:drug therapy / Q000201:enzymology; D006427:Hemin / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007908:Lens, Crystalline / Q000201:enzymology*; D009249:NADP / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism*; D010544:Peroxidases / Q000378:metabolism*; D010545:Peroxides / Q000378:metabolism; D010775:Photic Stimulation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling; D013481:Superoxides / Q000378:metabolism*; D020122:tert-Butylhydroperoxide
D009842:Oligopeptides; D010545:Peroxides; D013481:Superoxides; C013994:ferri-heme undecapeptide; D009249:NADP; D006427:Hemin; D020122:tert-Butylhydroperoxide; D006861:Hydrogen Peroxide; D010544:Peroxidases